Acta Veterinaria et Zootechnica Sinica

Cloning and Sequence Analysis of cDNA Encoding Porcine Tolllike Receptor 7

LI Qiang;LI Xue-wei;ZHU Li;CHEN Lei;LI Ming-zhou

2008, 39(5): 531-535. doi:

Abstract ( 1737 )

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In this study we cloned cDNA of Toll-like receptor 7(TLR7) gene from total RNA of mesenteric lymph nodes(MLN) in Neijiang pig by RT-PCR and RACE (rapid amplification of cDNA ends) . Sequence analysis indicated that the porcine TLR7 cDNA cloned was 3 834 bp in length (GenBank accession number: EF469730) and the open reading frame encodes a deduced protein with 1 050 amino acids residues. The analysis of deduced amino acids sequence indicated that TLR7 is a typical type I transmembrane protein with a LRR-RI ectodomains and a TIR cytoplasmic domains, the common structural features of TLRs. The comparison of the deduced amino acids sequence of porcine TLR7 with those of cattle, dog, human, cat and mouse showed that the amino acids homology similarity were 90.8%, 87.4%, 84.9%, 86.7% and 78.2%, respectively and the TLR7 is highly conserved among the different mammal species.

The Association Analysis between RBP4 Gene and ReproductionTraits in Beijing Black Pig

LUORENG Zhuo-ma;;WANG Li-xian;SUN Shi-duo

2008, 39(5): 536-539. doi:

Abstract ( 1435 )

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RBP4 gene was regarded as a candidate gene for reproduction traits of Beijing Black pig in this study, and the polymorphism was detected by PCRRFLP with restriction endomuclease MspⅠ. Three genotypes were found in Beijing Black pig, and the frequencies of allele A and B, polymorphism information content and heterozygosity were 0.529 6, 0.470 4, 0.374 1 and 0.498 2, respectively, which revealed a moderate polymorphism in this site. Statistical results of χ2 test indicated that this site fitted Hardy-Weinberg equilibrium (P>0.05). Meanwhile, the effect of polymorphism of RBP4 gene on TNB, NBA and WB were analyzed, the results indicated that the TNB, NBA and WB of multiparous sows with either genotype showed no significant difference (P>0.05). The sows of first parity with genotype BB had 1.04 and 0.31 litter size of TNB more than those with genotypes AA and AB, and the pigs of first parity with genotypes BB and AB had 1.47 and 1.35 litter size of NBA more than those with genotype AA, respectively. The results of genotype effective analysis indicated that allele B had the positive effect on TNB, NBA and WB, which might increase 0.282 0, 0.408 5 and 0.000 9, respectively, that was to say, allele B might be a favourable allele on TNB, NBA and WB.

Molecular Cloning of Bovine α-actinin1 Gene and the Association of Its Polymorphism with the Reproduction Traits

HUAI Ya-hong;;XU Shang-zhong;;WANG Shu-hui;GAO Xue;CHEN Jin-bao;REN Hong-yan

2008, 39(5): 540-546. doi:

Abstract ( 1522 )

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cDNA of α-actinin1 gene was cloned by RT-PCR based on the total RNA from bovine testicle. Furthermore, the sequence of cDNA was analysed using bioinformatics. A1-bd pair of primers and β-actin pair of primers were used to amplify α-actinin1 gene based on the total RNA from 12 bovine tissues, including liver, testicle, lung, rumen, uterus, small intestine, spleen, heart, fat, ovary, kidney and muscle. In addition, mutation was detected by DNA sequencing and PCR-RFLP in twinning Luxi cattle, Luxi cattle, Nili-Ravi bufflo, Murrah buffalo, Jinnan cattle, Holstein, Simmental and Nanyang cattle. The result indicated that the cDNA sequence was 2 911 bp, which contained an open reading frame of 2 679 bases (from 53 to 2 731 bp) which encoded an 892 amino acid. The α-actinin 1 protein had 3 mainly structural domains including CH, SPEC and EFh. α-actinin1 gene was expressed in liver, testicle, lung, rumen, uterus, small intestine, spleen, heart, fat and kidney, there was no express in ovary and muscle. Mutation of A/G was detected in intron 13 of α-actinin1 gene by 1.5% gelose gel electrophoresis. The mutation caused Hae Ⅲ polymorphism in group, so there were AA and AG genotypes. The association analysis of this Hae Ⅲ polymorphism with reproduction traits showed that Hae Ⅲ PCR restriction fragment length polymorphism (RFLP) genotypes may be no association with the number of calving (P＞0.05). In conclusion, the cDNA of α-actinin1 gene was obtained. A mutation was detected in all populations besides Holstein. There was no association between the number of calving and genotypes.

Cloning of Bovine Nanog Gene and Its Expression in Skin Fibroblast Cells

ZHENG Xi-bang;YUN Yan;HU Yong-ce;LI Yong;WANG Hua-yan;DOU Zhong-ying

2008, 39(5): 547-554. doi:

Abstract ( 1564 )

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The present study is to construct a eukaryotic expression vector of bovine Nanog, transfecting it into skin fibroblast, and further to obtain stable Nanog- transfected cell line, which can be used in nuclear transfer. By means of reverse transcription polymerase chain reaction (RT-PCR), cDNA of Nanog was cloned from tissues of fetal bovine primodial genital ridges at the age of about six weeks. It was inserted into pMD18-T vector, then into vector pCDNA3-FLAG by gene recombination technique. After being confirmed by restriction enzyme digestion and sequencing, the recombinant plasmid FLAG-Nanog was transfected into skin fibroblast cells. Selecting for two months with neomycine (G418), we obtained a stably transfected cell line. RT-PCR and Western Blotting were respectively used to detect the expression of Nanog mRNA and that of FLAG-Nanog fusion protein. In addition, immunostaining assay was performed to investigate whether the cell line is characteristic with stem cells. The results showed that (1) complete cording sequences (CDS) of Nanog was cloned from fetal bovine primodial genital ridges; (2) its eukaryotic expression vector was constructed, which expressed efficiently in the skin fibroblast cells; (3)Stage Specific Embryonic Antigen 4 (SSEA-4), factor Oct-4 as well as Nanog, required to maintain pluripotency of ES cells, was readily detectable in these Nanog-expressing fibroblast cells, indicating that the cell line is to some extent pluripotent. Our present study will lay a good foundation for further studying functions of Nanog, especially its role on early embryo development, production of transgenic animals, and derivation of embryonic stem cell lines from farm animals.

Genetic Diversity of Microsatellite DNA among Ten Sheep Breeds

ZHONG Tao;MA Yue-hui;GUAN Wei-jun;LING Ying-hui;GUO Jun;ZHAO Qian-jun;HE Xiao-hong

2008, 39(5): 555-561. doi:

Abstract ( 1370 )

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In order to investigate the genetic diversity of Chinese indigenous sheep breeds, genetic variation of 9 Chinese indigenous sheep breeds and one introduced breed were examined using 21 microsatellite DNA markers and fluorescence PCR. All the 21 markers are highly polymorphic. The polymorphism information content (PIC) and genetic heterozygosis showed that the Chinese local breeds have high genetic diversity. A total of 342 alleles were detected, and the effective allele number ranged from 2.175 2 to 9.499 7. The average heterozygosity of loci varied from 0.524 8 to 0.855 1, and the average heterozygosity of breeds were from 0.633 to 0.761. The phylogenetic trees(NJ and UPGMA) analysis were generally in accordance with their origins, breeding history and localities.

Genetic Differentiation and Gene Flow among Seven Indigenous Goat Populations in South of China

YANG Zhang-ping;MAO Yong-jiang;MA Yue-hui;WANG Zhi-guo;WANG Qing-hua;CHANG Hong;CHANG Guo-bin;SUN Wei;LI Shu-chun

2008, 39(5): 562-569. doi:

Abstract ( 1583 )

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The genetic differentiation, gene flow, the relationship between geographical distance and genetic differentiation among seven indigenous goat populations in south of China were estimated using 23 microsatellites. The trees were constructed from DC distances and the data were analyzed by STRUCTURE (Ver 2.0). The global heterozygote excess across all populations (Fit) amounted to －7.73% (P<0.001). The overall significant (P<0.001) excess of heterozygotes within populations amounted to －26.5%. The 7 goat populations were highly differentiated (Fst= 14.84%, P<0.001) at all loci, which indicated that 14.84% of the total genetic variation came from breed differences, and the remaining 85.16% came from differences among individuals in each population. The average number of effective migrants exchanged per generation (Nem) ranged from 0.831 3 (Yichang White goat and Huanghuai goat) to 3.410 3（Matou goat and Xiangdong Balck goat）, and the average value was 1.577 0. No significant positive relationships between the level of genetic differentiation and geographical distance, genetic distances among 7 goat populations were detected. With the application of prior population information, cluster analysis by STRUCTURE (Ver. 2.0) achieved posterior probabilities 99.1%, 98%, 96.2%, 96.6%, 98.7%, 98.7% and 98.7% of correctly assigning sampling populations to different inferred clusters for Yichang White goat, Matou goat, Xiangdong Balck goat, Fuqing goat, Daiyun goat, Huanghuai goat and Yangtse River Delta White goat, respectively. The inferred clusters by STRUCTURE were similar with the trees constructed from DC distances. From the results it indicated that the reason for the genetic differentiation among seven indigenous goat populations in south of China is natural selection.

Genetic Polymorphism and Cluster Analysis on Blood Protein (Enzyme) of Mongolian Horses

YANG Hong;MENG Ke;WANG Ye-hui;MANG Lai

2008, 39(5): 570-575. doi:

Abstract ( 982 )

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Eight types of blood proteins in serum samples from 221 Mongolian horses (including Wuzhumuqin horse, Wushen horse, Xinihe horse and Baerhu horse) were detected on polymorphism by vertical plate discontinuous polyacrylamide gel electrophoresis (PAGE). Genotype frequencies, allele frequency and genetic heterozygosity of these sites were calculated and the relationship and the genetic distance between the groups were discussed through cluster analysis.The results indicated that in addition α1-BGlycoprotein（A1B）was of monomorphism, the remaining 7 blood protein (enzyme) sites were of polymorphism in varying ways. The statistical results showed that the average heterozygosity (H) of these eight protein (enzyme) sites in Wuzhumuqin horse, Wushen horse, Xinihe horse and Baerhu horse were 0.329 6, 0.305 6, 0.378 3, 0.366 6, respectively. The cluster analysis suggested that the genetic distance between Baerhu horse and Xinihe horse was closer, however, the genetic distance between these two types of horses and the other two breeds were relatively farther.

Canonical Correlation Analysis of Traits for Eleven Native Chicken Breeds

WU Xin-sheng;XU Qi;XIAO Xiao-jun;ZHANG Xue-yu;WU Sheng-long;BAO Wen-bin;LI Bi-chun;CHEN Guo-hong

2008, 39(5): 576-581. doi:

Abstract ( 1004 )

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Seventeen variants of four sets of traits including body measurement traits, carcass traits, egg laying traits and ecological traits were analysed using canonical correlation analysis in Chinese native chicken breeds. The results showed that highly significant correlation coefficients of the first canonical correlation were obtained between body measurement traits and carcass traits（0.998）,body measurement traits and ecological traits（0.997）, as well as between carcass traits and ecological traits（0.998）. They accounted for 38.17%, 42.73% and 42.13% of total correlation, respectively. The first canonical correlation coefficient between body measurement and egg laying traits（0.965）, as well as between egg laying traits and ecological traits（0.984）were significant. They a ccounted for 42.55% and 66.79% of total correlation, respectively. The results of analysis may be useful for genetics and breeding of chickens.

Cloning of MSTN Gene and Research of Relationship between MSTN Gene Expression and Different Energy Diets or IGFI,GH in Serum of Xupu Geese

HU Jun-peng;HE Rui-guo;FAN Wei-xing;CAO Ai-qing

2008, 39(5): 582-587. doi:

Abstract ( 925 )

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Genome RNA was extracted from leg muscle of Xupu geese,MSTN gene mRNA was amplified by using RT-PCR, the PCR product was cloned into pGEM-T vector.The MSTN gene was isolated and sequenced from the positive clones screened based on the MSTN gene clone, using β-actin as inner control, constructing an optimal semi-quantitative RT-PCR method, the difference of MSTN gene expression at 21 and 70 days by feeding different energy diets, at one time we detected IGF-I and GH in serum. The unique DNA fragment about 1 128 bp in length was obtained. Sequence analysis revealed that it has a length of 1 128 bp which encodes a 375 amino peptide. Compared with homologous sequence of chicken, duck and Landes geese, it displayed a fairly high conservation. The homologue of the nucleotide sequence are 94%,94%,99%, respectively; the homologue of the amino acid sequence deduced are 98%, 97%,98%, respectively. The results showed the expressions of MSTN in Xupu geese were showed no significantly differences at 21 days; but the expression of MSTN was C>B>A by 70 days; During 2170 days period, the expression of MSTN was changed with IGF-I in serum; it was not associated with GH in serum. The different energy influenced MSTN gene expression of Xupu geese after 21 days. The present study laid a foundation for further investigating MSTN gene in waterfowl.

Effects of EGF，IGF-I on in vitro Maturation and Cleavage of Ovine Oocytes

LIU Chou-sheng;LU Hui-ning;ZHANG Li-ping ;WANG Zhi-gang;ZHAO Jun-jin ;MENG Fei

2008, 39(5): 588-593. doi:

Abstract ( 907 )

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To establish an optimal conditions for in vitro maturation of ovine oocytes, we studied the effects of EGF, IGF-I and the union use of both of them on in vitro maturation and cleavage of ovine oocytes. The results showed that the maturation and cleavage rates derived from oocytes matured in medium supplemented with 50 ng/mL EGF alone were 71.2% and 45.5%， respectively, which was significantly higher than those in control and those with 10，20，30，40 ng/mL EGF, and the highest maturation and cleavage rates present in medium with 100 ng/mL EGF, were 70.7% and 58.5%, respectively. While the oocytes were only treated with 40 ng/mL IGFI, the maturation and cleavage rates were 70.7% and 58.5%, respectively, which was noticeably higher than that of the control and those with 10，20， 60，80，100 ng/mL IGF-I, but the maturation and cleavage rates were only 38.8% and 20.0%, respectively when the concentration of IGF-I was 100 ng/mL, the difference is remarkable compared to the experimental groups and control group. When treated by the combination of 50 ng/mL EGF and 40 ng/mL IGF-I, the maturation and cleavage rates were 85.6% and 61.0%, respectively, which is significantly higher than the single use of 50 ng/mL EGF or 40 ng/mL IGF-I and the control group.

Effeccts of Casein Hydrolysate on the Intestinal Growth and Development of IUGR Piglets

ZHAO Nian;ZHANG Li-li;JIN Rong;DU Wei;WANG Tian

2008, 39(5): 594-400. doi:

Abstract ( 931 )

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The effects of casein hydrolysate on the growth and development of intestine in intrauterine growth restriction (IUGR) piglets were examined in this study Fifteen piglets were selected and divided into newborn control group (N), casein group (C) and casein hydrolysate group (CH). The cow-milk was taken as the basic feed, piglets in group C and CH were fed milk substituted casein solution and casein hydrolysate solution for 10% milk, respectively. The piglets were slaughtered and sampled 3 days after feeding. The intestinal length of piglets in group CH was significantly higher than that in group C (P<0.05). The villus height/crypt depth of distal jejunum and primal ileum in group CH was significantly higher than that in group C (P<0.05). The protein and DNA contents in intestinal mocosa in group CH were significantly higher than those in group C (P<0.05). The DNA concentration in intestinal mocosa in group CH was higher than that in group C (P=0.056). The activity and relative activity of maltase in intestinal mucosa in group CH were significantly higher than those in group C and N (P<0.05 or P<0.01). The activity of AKP in intestinal mucosa in group CH was significantly higher than that in group C (P<0.05). These results suggested that casein hydrolysate can stimulate the hyperplasia of epithelium cell and increase the activities of maltase and AKP in small intestinal mucosa.

Study on the Variation of Amylase, Trypsin and Chymotrypsin Activities in Jejunal Fluid of Peking Duck

ZHAO Feng;ZHANG Hong-fu;HOU Shui-sheng;ZHANG Zi-yi

2008, 39(5): 601-607. doi:

Abstract ( 928 )

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The variation of jejunal digesta flux in cannula and digestive enzymes activities in jejunal fluid of Peking duck were investigated to study the scheme of representive jejunal fluid sample. The 3×6 factorial treatment was performed in two experiments and jejunal digesta samples were collected for 1 h out of every 4 h on d 16, 18 and 20 of the experiment. In experiment 1, fifteen male Peking ducks of 18 wk of age fitted a T type jejunal cannula were randomly allotted into 5 replication with 3 birds per replication. The volume per h of jejunal digesta in cannula was determined. In experiment 2, twenty-four cannulated ducks were randomly selected and divided into 6 replication with 4 birds per replication. The amylase, trypsin and chymotrypsin activities in jejunal fluid were determined. The results showed that there was obvious variation of digesta flux in cannula with collection time and collection day. Collection time significantly affected the average activities of amylase, trypsin and chymotrypsin in 24 h cycle (P＜0.01). Significant differences were also found in the average activities of amylase and chymotrypsin among days (P<0.05). Three digestive enzymes activities were declining with collection time in a 24 h cycle. According to the results, it is possible to develop a representative sampling scheme that sampling for 1 h out of every 4 h in the daylight at 1 day interval for 5 days for studying in endogenous digestive enzymes.

Effects of Lipopolysaccharide Challenge on PPARγ mRNA Expression in Immune Cells and Immune Organs of Weanling Piglet

SHI Jun-xia;LIU Yu-lan;LU Jing;FAN Wei;ZHAO Sheng-jun;ZHU Hui-ling;HOU Yong-qing;DING Bin-ying;GUO Guang-lun

2008, 39(5): 608-613. doi:

Abstract ( 950 )

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The experiment was conducted to investigate the effects of lipopolysaccharide (LPS) on PPARγ mRNA expression in the peripheral white blood cells, thymus, spleen and intestinal lymphoid node of weanling piglets. 12 pigs were divided into two groups (6 replicates/treatment and 1 pig/replicate). The piglets in the treatment group were injected intraperitoneally with 100 μg/kg BW LPS, whereas pigs in the control group were injected with an equivalent amount of sterile saline. Blood samples were collected at 1.5 and 3 h post-challenge for isolating plasma or white blood cells. Following blood collection at 3 h, the piglets were slaughtered to collect thymus, spleens and intestinal lymph node for detection. The results showed that: (1) Tumour necrosis factor-α and cortisol were increased significantly 1.5 and 3 h after LPS challenge (P<0.001), insulin was decreased significantly (P<0.001), and glucogan was increased significantly (P<0.001) 3 h after LPS challenge. Additionally, LPS challenge changed blood biochemical parameters significantly. These results indicate that LPS challenge induces acute immunological stress in piglets. (2)LPS challenge increased PPARγ mRNA expression in the peripheral white blood cells (P<0.001), thymus (P<0.05) and intestinal lymphoid node (P<0.05), which indicates that immunological stress induced PPARγ mRNA expression of immune cells and immune organs. These results suggest that PPARγ may play an important role in regulation of immunological stress in pigs, and may be a new target for alleviating immunological stress of piglets.

The Homology Molecular Identification of Airborne Escherichia coli Isolated from Indoor and Outdoor Air of Chicken Houses

DUAN Hui-yong;CHAI Tong-jie

2008, 39(5): 614-620. doi:

Abstract ( 1242 )

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In order to study the transmission of E. coli from chicken houses to their ambient air, the air samples, including indoor air, upwind air and downwind air of 5 chicken houses were collected using ANDERSEN-6 stages sampler and RCS. Chicken feces samples were collected according to the standard method. E. coli strains were isolated from these samples . The enterobacterial repetitive intergenic consensus (ERIC)-PCR method was applied to generate genomic amplification products of isolated E. coli. According to the concentration and genetic similarity of E. coli in every sampling site, the transmission of bioaerosol from animal houses to their ambient can be identified. The results showed that the culturable E. coli concentration in indoor air in 5 chicken houses were higher than that in upwind and downwind (P<0.05 or P<0.01), but there were no significant differences in E. coli concentration among different downwind site (P>0.05). The fingerprints and the phylogenetic tree indicated that a part of the E. coli (34.1%) isolated from indoor air had the same ERIC-PCR fingerprints with those isolated from feces, the most of E. coli (54.5%) isolated from downwind 10 m, 50 m, 100 m even 200 m away from the houses had the same ERICPCR fingerprints with those isolated from indoor air or feces. But E.coli isolated from upwind air had the low similarity coefficients (73%-92%) with those isolated from indoor air or feces. So, it was concluded that the E. coli in chicken feces can be aerosolized and transmitted into the indoor and outdoor air, especially the downwind air. This study revealed the transmitting rule of airborne E. coli bioaerosol. Hygienic measures took in animal farms have the significance of public hygiene and epidemiology.

Insertion Mutagenesis in Salmonella pullorum by Mini-Tn5 Transposon

GENG Shi-zhong;JIAO Xin-an;CHEN Xiao-juan;PAN Zhi-ming;ZHANG Hui;CHEN Xiang

2008, 39(5): 621-626. doi:

Abstract ( 1075 )

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CmR and KmR genes were transposed into genomic DNA of Salmonella pullorum to produce a library of mutants of Salmonella pullorum by pUTminiTn5Km2(Cm) which was constructed by inserting CmR gene into pUTminiTn5Km2. Then some defective mutants were screened. Mutants were identified by basic characteristic of Salmonella pullorum, special PCR and gene location. The results showed that CmR and KmR genes were integrated into genome of Salmonella pullorum mutants and each mutant has one and only one different locus of transposon in the genome. These results provided essential base for studying the function gene of Salmonella pullorum and screening special mutants.

Nucleotide Sequence and Phylogenetic Analysis of LIPI-1 Gene Cluster from Listeria monocytogenes

TAN Bing-qian;;HE Qi-gai;XIAO Jun;CHEN Huan-chun

2008, 39(5): 627-633. doi:

Abstract ( 853 )

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The LIPI-1 gene cluster of Listeria monocytogenes isolate strain XFL0605 was segmentally amplified by PCR and sequenced. The results showed that the fragment was 8 558 bp，and containing all the members of the prfAregulated virulence gene cluster such as prfA, hly, plcA, plcB, mpl and actA. Phylogenetic analysis of the isolate XFL0605 and other reference L. monocytogenes, L. seeligeri and L. ivanovii strains based on the gene sequence demonstrated clearly that the evolutionary relationship of strains were different. It revealed that recombination of LIPI-1 has different assembly process. Furthermore, we predicted the signal peptides and transmember domain and their cleavage positions, and investigated the PrfA DNA binding sites of the LIPI-1 virulence genes. The hlyA gene deduced amino acid of XFL0605 contains a PEST-like sequence at the N-terminus, and a conserved undecapeptide, ECTGLAWEWWR, at the C terminus. It also showed that actA gene of XFL0605 encode 604 amino acids but 105 bp nucleotide missing in proline-rich region, only contains two E/DFPPPPXD/E repeat sequences.

Construction and Characteristics of an ApxIV Gene Mutant of Actinobacillus pleuropneumoniae Serotype 7

LIU Jin-lin;GUO Yi;TAN Chen;CHEN Yan;BEI Wei-cheng;CHEN Huan-chun;

2008, 39(5): 634-638. doi:

Abstract ( 984 )

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In order to study pathogenicity of the fourth RTX toxin ApxIV in the infection of Actinobacillus pleuropneumoniae, an Actinobacillus pleuropneumoniae apxIVA mutant was constructed by transconjugation and counterselection method,using an A. pleuropneumoniae HB04 strain (serotype 7) as parental strain. The apxIV mutant was confirmed by PCR, sequence analysis, Southern blot and heredity stability assays. The biological characteristics of this mutant strain was further investigated. The mutant possesses the same growth rate as its parental strain, but it was less virulent on mice. These results indicated that apxIV was an important virulence factor in the pathogenesis of A. pleuropneumoniae infection. This study have a further illumination on the pathogenicity of A. pleuropneumoniae and it will facilitate the development of more reasonable live attenuated vaccines of APP.

Preparation and Immune Efficacy of Inactivated Vaccine against Porcine Circovirus 2

DONG Xin-tian;LI Yu-feng;JIANG Ping;WANG Xian-wei;FENG Zhi-xin;YU Li-li

2008, 39(5): 639-644. doi:

Abstract ( 984 )

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Porcine circovirus 2 was inactivated with 0.2% formaldehyde under 37℃, then was passaged on PK-15 cell three times and the inactivation effect was estimated by PCR method. The immune efficacies of the oil-adjuvant and the aluminum hydroxide-adjuvant inactivated vaccines against PCV2 were evaluated in mice. Mice were vaccinated subcutaneously twice at a threeweek interval. The specific ELISA antibody against PCV2 were detected at the 21st day after the first immunization and at the 14th day after the boost. In order to compare the immune efficacy between the homemade oilbased and imported oil-based vaccines, piglets were immunized with two inactivated PCV2 vaccines and the immune efficay was evaluated by presence or absence of seroconversion, clinical signs (fever), gross lesions, growth parameters and viremia. The results were as follows: ① PCV2 was inactivated completely with 0.2% formaldehyde after incubation for 16 h. ② The good immune response could be induced by two vaccines in mice and the immune efficacy of the oil-adjuvant inactivated vaccine was higher than that of the aluminum hydroxide-adjuvant inactivated vaccine. ③ High ELISA antibody titers of two oil-adjuvant vaccines could be detected in piglets after the boost. After challenge the rectal temperatures of all piglets were recorded. Average temperature of piglets in challenge control group was higher than that of immunized piglets and no pyrexia was observed in mock piglets. The average weight was increased in two immunized groups and mock group, but decreased in challenge control group. Comparison of average daily weight growth (ADWG) of vaccinated pigs with that of mock pigs revealed statistically significant difference between them (P<0.05). The fact that two vaccines could reduce the persistence time of PCV2 in piglets was indicated by detection of viremia and PCV2 in different tissues of challenged piglets. It could be concluded that inactivated PCV2 vaccines could induce immune response in mice and piglets and could protect piglets from PCV2 infection.

Distribution of Pseudorabies Virus in Tissues of Latently Infected Pigs

LONG Xiao-ting;LIU Jun-lei;GUO Hong;XI Yang;QIU De-xin;CHEN Huan-chun;ZHOU Rui

2008, 39(5): 645-651. doi:

Abstract ( 1636 )

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Latent infection is one of the most important obstacles for prevention, control and eradication of pseudorabies in the swine industry. In this study, a pig pseudorabies virus (PRV)-latency model was established by vaccinating PRVfree piglets with commercial PRV inactivated vaccine, followed by challenge with a genetically marked PRV strain gG-/LacZ⁺.Then the latency was reactivated by administration of dexamethasone. The distribution of PRV in the tissues of latently infected and re-activated pigs was studied by immunohistochemistry SABC staining with antibodies against β-galactosidase. The results showed that β-galactosidase positive cells were mainly found in the nerve tissues (cerebrum, cerebellum, brain stem, trigeminal nerve and optic nerve) and other tissues (tonsil, lung and kidney), and the quantities of positive cells decreased along the time post gG-/LacZ+ infection. After administration of dexamethasone the numbers of positive cells increased in these tissues.

Protection of Lycium Barbarum Polysaccharides on Diethylstilbestrol Induced Oxidative Damage in Testis of Hamster

MA Ai-tuan;CHEN Yao-xing;WANG Zi-xu;DONG Yu-lan

2008, 39(5): 652-657. doi:

Abstract ( 960 )

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Protective effects of Lycium Barbarum polysaccharides (LBP) on diethylstilbestrol (DES) induced oxidative damage in hamster testis was studied in this experiment. Adult hamsters (Mesocricetus auratus) were randomly divided into 4 groups: Control group, DES group, DES+LBP group, DES+V_C+V_E group. Absolute and relative testicular weight were calculated, morphological changes of testis were observed and the contents of SOD, GSH-Px, T-AOC, MDA in testis tissue and plasma were detected after 7 successive days treatment. The results showed that LBP had an evident effect in reversing the reproductive damage induced by DES. In DES+LBP group, the absolute and relative testicular weight were significantly upgraded (P <0.05) and the seminiferous tubules were lined up in order. Levels of SOD and TAOC elevated markedly in LBP group (P <0.01),which was similar with control, but MDA showed the reverse tendency. There was no significant difference between LBP and vitamin group. It is concluded that LBP could efficiently restrain the production of reactive oxygen species, inhibit the generation of lipid peroxidation, accordingly attenuate the reproductive toxicity of DES. These results indicated that LBP is one of the active components in attenuating the oxidative damage induced by environmental estrogens.

Effect of Dietary High Copper on the Bursa of Fabricius in the Chicken

XU Zhi-yong;CUI Heng-min;PENG Xi;ZHU Kui-cheng;DENG Jun-liang;WU Qiang;YANG Fan;ZHAO Li

2008, 39(5): 658-664. doi:

Abstract ( 1010 )

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The experiment was conducted with the objective of examining the effects of high copper on the bursa of Fabricius in the chicken by the methods of experimental pathology and flow cytometry (FCM). 420 one-day-old Avian chickens were randomly divided into seven groups, and fed on control diet （Cu 11 mg/kg） and high copper diets （Cu 100 mg/kg,high copper group Ⅰ；Cu 200 mg/kg,high copper group Ⅱ；Cu 300 mg/kg,high copper group Ⅲ；Cu 400 mg/kg,high copper group Ⅳ；Cu 500 mg/kg, high copper group Ⅴ；Cu 600 mg/kg, high copper group Ⅵ）for six weeks,respectively. The results showed that histological changes were observed in high copper groups Ⅲ，Ⅳ，Ⅴand Ⅵ in comparison with those of control group. The weight and the growth index of the bursa of Fabricius were higher in high copper groupsⅠand Ⅱ（P＞0.05）,and lower in high copper groups Ⅲ，Ⅳ，Ⅴand Ⅵ than that in control group（P＜0.05 or P＜0.01）. The proliferation index（PI） of the bursa of Fabricius was increased in high copper groups Ⅰand Ⅱ（P＞0.05）,and decreased in high copper groups Ⅲ，Ⅳ，Ⅴand Ⅵ（P＜0.05 or P＜0.01）compared with that of control group. Percentage of apoptotic lymphocytes in the bursa of Fabricius was higher in high copper groups Ⅲ，Ⅳ，Ⅴand Ⅵ than that in control group（P＜0.05 or P＜0.01）.It is concluded that dietary copper in excess of 300 mg/kg inhibited the development of bursa of Fabricius，caused pathological changes in the bursa of Fabricius and impaired humoral immune function，and that 11200 mg/kg of copper in diet was helpful for the development of the bursa of Fabricius.

Effect of Verapamil on Pulmonary Hypertension Syndrome via Mast Cells of Lung and Heart in Broiler Chickens

OU De-yuan;QIAO Jian;ZHANG Jian-jun;SUN Mao-hong;DONG Shi-shan

2008, 39(5): 665-671. doi:

Abstract ( 902 )

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The effect of Ca²⁺ channel of mast cells (MC) on the development of pulmonary hypertension syndrome (PHS) was investigated in broiler chickens. The birds with PHS were induced by low ambient temperature. The pulmonary artery pressure was measured using right cardiac catheter. MC were stained with Alicience blue. Histamine was shown using immunohistochemistry. The deposit of Ca²⁺ was observed in MC by transmission electron micrograph. The results showed: (1) The Ca²⁺ channel blocking agent (verapamil) can decrease the pulmonary artery systolic pressure and pulmonary artery diastolic pressure significantly. (2) Verapamil decreased the right ventricular pressure and ascites heart index（P＜0.05）. (3) Verapamil promoted the number of MC in peripheral pulmonary blood vessel walls and right ventricle（P＜0.05）, and markedly decreased（P＜0.05） the ratio of degranulation of MC. (4) Verapamil increased the number of histamine positive cells in the lung（P＜0.05）. (5) Intracellular Ca²⁺ was increased significantly in degraunlated MC. These results suggested that verapamil inhibited the degranulation of MC and releasing mediators to prevent the development of the PHS.

Construction, Expression and Immunization of DNA Vaccine pcISI Harboring Two Copies of INHα1-32

CAO Shao-xian;;XING Zhao-fang;ZHANG De-kun;SHU Deng-qun;YANG Li-guo;

2008, 39(5): 672-676. doi:

Abstract ( 1562 )

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To construct an inhinbin (INH) expression vector with high immunogenicity, INHα1-32 gene was incorporated into the S gene of pcIS between amino acid residues 112 and 113. The resultant plasmid pcISI with two copies of INHα_1-32 was identified by restriction endonuclease digestion and sequencing. The plasmid pcISI was then introduced into HeLa cells via liposome-mediated transfection and the immunogenicity of recombinant protein was detected by ELISA. The results showed that the recombinant gene expressed in HeLa cells, and the INH immunogenicity of ISI fusion protein was higher than that of IS fusion protein, indicating the ISI expression vector was constructed successfully. Five of six rats immunized with pcISI produced antibodies against INH in vivo. In addition, the P/N values of antibodies were higher than those immunized with pcIS from 2nd week to 6th week. In conclusion, the constructed pcISI could express INH with high immunogenicity.

Selenium Supplementation on Growth Performance and Immune Function of Oxidative Stressed Piglet

YUAN Shi-bin;YU Bing;CHEN Dai-wen

2008, 39(5): 677-681. doi:

Abstract ( 1360 )

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Effect of selenium supplementation on growth performance and immune function of oxidative stressed piglets were studied in this experiment.Fortyeight weanling piglets without vaccination for swine fever were allotted into four groups according to a one factorial design.Each group had six replications with two piglets each.Piglets received injection of diquat at the beginning of the trial.Selenium was supplemented at 0,0.2,0.4 or 0.6 mg/kg in the diets.At the end of the third week of the trail,all the pigs were injected with swine fever vaccine.The results showed that selenium supplementation could alleviate the impacts of oxidative stress.The growth performance and immune function of oxidative stressed piglets were significantly improved as selenium level in the diet increased,compared with other groups,the effect of 0.6 mg/kg group was the best.

Comparative Study on Immunological Activity Survival of SoybeanAgglutinin in Digestive Tracts of Different Species of Animals

WANG Li-min; HU Hai-xia;WANG Tao;QIN Gui-xin

2008, 39(5): 682-686. doi:

Abstract ( 1309 )

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In this experiment, different animals (sheep as ruminants, dog as carnivores, rabbit as rodents, pig as omnivores and chicken as fowls) were used to compare the immunological activity survival disciplinarian of SBA in the chyme within different parts of digestive tracts. Each animal aged the half of their sexual maturity, 8 each species with similar conditions (sex) were looked as a repeat and fed the diet containing 20% raw soybean and 1%Cr2O3 ( as exogenous indicator). Two hours after the last meal of two weeks' feeding, the chymes in duodenum, upper-jejunum, middle-jejunum, lower-jejunum, ileum, appendix and colon were taken. The SBA having immunological activity in the chymes and feeds were detected with ELISA and the Cr in the chymes and feeds were detected with the absorption spectroscopy. The survival rates of SBA in different parts of digestive tracts in the same animal were different and showed the downtrend from duodenum to colon, however, the SBA were existent in all of the parts of digestive tracts. The changes of the survival rate from duodenum to front-jejunum were significant（P＜0.05）in all of the animals. There were different survival rates in the same parts of digestive tracts in the different animals. Rabbits possesed the lowest survival rates in all of the digestive parts. Sheep and chickens were higher than pigs and dogs from duodenum to front-jejunum. Behind jejunum, pigs, sheep and chicken had more survival rates than that of others. There are different immunological activity survival disciplinarian of SBA in different animals and their different parts of intestinal tracts. The antinutritional mechanism and effects of SBA on different species of animals maybe different.

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